Hello Azam Mahmoudi Aznaveh

You could refer to the research article below which could be helpful.

Summary:

The mycotoxin 2-Amino-14,16-dimethyloctadecan-3-ol (AOD) has been isolated from cultures of the fungus Fusarium avenaceum, one of the most prevalent Fusarium species. AOD is an analogue of sphinganine and 1-deoxysphinganine, important intermediates in the de novo biosynthesis of cellular sphingolipids. Here we studied cellular effects of AOD using the human liver cell line HepG2 as a model system. AOD (10 μM) induced a transient accumulation of vacuoles in the cells. The effect was observed at non-cytotoxic concentrations and was not linked to cell death processes. Proteomic analyses indicated that protein degradation and/or vesicular transport may be a target for AOD. Further studies revealed that AOD had only minor effects on the initiation rate of macropinocytosis and autophagy. However, the AOD-induced vacuoles were lysosomal-associated membrane protein-1 (LAMP-1) positive, suggesting that they most likely originate from lysosomes or late endosomes. Accordingly, both endosomal and autophagic protein degradation were inhibited. Further studies revealed that treatment with concanamycin A or chloroquine completely blocked the AOD-induced vacuolization, suggesting that the vacuolization is dependent of acidic lysosomes. Overall, the results strongly suggest that the increased vacuolization is due to an accumulation of AOD in lysosomes or late endosomes thereby disturbing the later stages of the endolysosomal process.

Research Article:

Article The Fusarium mycotoxin, 2-Amino-14,16-dimethyloctadecan-3-ol...

Best Wishes.

Dear Malcolm Nobre,

Thanks for the useful data. I will consider that.

I think the cells are under stressful condition for some reason and it will be detached from the surface. Try to maintain these cells on freshly prepared cell culture media and handle with care. All the best.

Considering your other cell lines are fine, I don't think there is any problem with handling or contamination. Try reverting to an older freeze. After a lot of passages, the cells will begin to lose their identity and start showing abnormalities and dying/clumping every now and then.